第一部分 : 防曬係數之體內及體外評估的相關性 , 第二部分 : 膠原蛋白膜的特性解析(Ⅱ)

First Part : The Correlation of In-Vivo and In-Vitro Measurement of SPF(Sun Protection Factor) , Second Part : Characterization of Collagen Film(Ⅱ)

林正雯(Lin Cheng-Wen)

指導教授 ： 許明照

臺北醫學大學/藥學院/藥學研究所/碩士(2000年)

摘要

第一部分: 本實驗的目的在於探討體內與體外之防曬係數評估的相關性，希望能夠用體外取代體內的評估方式，以節省人力和金錢，更有助於產品設計的方便性。本次實驗採用八個防曬樣品，包括兩種標準品[FDA(SPF=4)、COLIPA(SPF=15)]和六種市售商品。 在體內試驗方面，利用受測者背部的皮膚作為照射範圍，塗抹劑量是以每平方公分2毫克來計算，塗抹後等待15分鐘，再以光源於塗抹處進行照射；此光源是採用Multiport UV Solar Simulator，其含有六個光源輸出管，每管之間的劑量以25%遞增，經適量照射後24小時，由皮膚科醫生來判讀最小紅斑劑量(MED)。另一方面，體外試驗的過程中，則以3M transpore tape來當作人造皮，並利用紫外光穿透率分析儀(Ultraviolet Transmittance Analyzer UV-1000S)的Xenon燈來模擬光源，測試結果會由其附屬軟體自動計算出防曬係數值。 比較體外及體內試驗的結果發現，由體內試驗所計算出的防曬係數值與其標示值較相近，表示在此實驗中防曬係數值於體內試驗之測試結果應可信賴；然而利用統計方法(T-test/ANOVA)來分析體內和體外的相關性，發現在大部分產品中，兩種方法的防曬係數值具有顯著的差異性；因此，防曬係數值在體內和體外試驗的相關性目前仍無法建立；所以利用紫外光穿透率分析儀來評估防曬產品的防曬係數值在體內和體外的相關性之建立，或許不是一個實際可行的分析方法。 第二部分: 由於膠原蛋白具有生物相容性、生物降解性及熱安定等優點，所以通常利用膠原蛋白作成不同型式如海綿 (sponges)、膜 (films) 和微粒 (microparticles) 劑型作為藥物載體。膠原蛋白載體的特性可進一步利用交聯劑 (cross-linking agent) 加以修飾，以調控適用於不同藥物特質。本實驗中採用化學性的方法，以最常用的交聯劑-戊二醛 (Glutaraldehyde) 作為交聯劑。首先利用數字式黏度計測量不同戊二醛濃度 (0%，0.05%，0.075%，0.1%，0.2%，0.25%，0.3%)，以及助溶劑 Cremopher RH40 (0%，1%，5%，10%) 和 1% 活性藥物 (維生素A-Palmitate、C-Palmitate、E-Acetate、A-Acid) 對膠原蛋白的交聯速度與程度之影響。最後再以不同戊二醛濃度 (0%，0.05%，0.075%，0.1%，0.2%，0.25%，0.3%) 交聯製備膠原蛋白膜，而以溶液 (solution) 和膠體 (gel) 的藥物形式 (藥物同前) 評估藥物滲透之機制。 由實驗結果顯示膠原蛋白黏度會隨著戊二醛濃度之增加而提高，但當戊二醛濃度達到0.2% 以上時，其黏度會到達平衡而成一定值。另一方面，添加Cremopher RH40時，膠原蛋白黏度反而會隨著Cremopher RH40濃度之增加而降低，但活性藥物卻並不會影響膠原蛋白的黏度。在滲透實驗中，發現大部分藥物溶液或膠體在不同戊二醛濃度交聯的膠原蛋白膜中，滲透通量 (Flux) 皆會隨著戊二醛濃度增加而降低，當戊二醛濃度達到0.2%以上時，藥物滲透通量會達到平衡；可是在維生素A酸藥物中不論是以溶液或膠體的藥物形式，戊二醛濃度達到0.2% 以上時，藥物滲透通量反而會有增加的趨勢，但由藥物對膠原蛋白膜之分配係數的差異仍無法說明此一現象。

關鍵字

防曬係數 ； 防曬劑 ； 膠原蛋白 ； 交鏈劑 ； 戊二醛

並列摘要

First Part： In this study, the correlation of measuring SPF value between in-vivo and in-vitro tests was investigated for the purpose of monitoring the in-vivo efficacy of sunscreen products by using in-vitro tests instead of in-vivo tests to lessen labor burden. Eight products, including two standards of FDA (SPF=4) and COLIPA (SPF=15) and six commercial products, were included for comparison. For the in-vivo test, the area for application of the products was delineated by using a template on the back of the volunteer. The quantity of test products was weighed and evenly distributed on the skin to have a concentration of 2 mg/cm2 using a finger stall. The lag time between application of products and UV irradiation was about 15 minutes. A Multiport UV Solar Simulator 601 with six outputs was employed as the UV source. Six test sites were exposed to UV irradiation in every treated area with a progression of UV doses of 25% from site to site. The minimal erythemal dose (MED) was assessed visually 24h after UV exposure. For the in-vitro tests, sunscreen products were evenly distributed on a 3M Transpore® tape (3M company) and the transmittance was measured by a UV Transmittance Analyzer equipped with a single-flash xenon lamp. The results were automatically transformed into a SPF value with a build-in program. The results demonstrated that the SPF values of in-vivo tests for each product was close to its claimed value, indicating the SPF values measured in this study was reliable. However, statistical analysis either by t-test or ANOVA showed that a significant difference of SPF value between in-vivo and in-vitro tests existed for most of products. Thus, a correlation of SPF measurement between in-vitro and in-vivo tests was not possibly constructed. According to these results, the in vitro testing using UV Transmittance Analyzer to measure SPF value was not a reliable in-vitro method to correlate with the results of in vivo tests and was not a practical way to monitor the effectiveness of sunscreen products. Second Part： Because of its biological properties and easy availability, collagen has been processed into several textures and shapes including sponges, films, membranes, and micro-particles as drug carrier for delivery. Collagen-based biomaterials can be further modified by crosslinking to match the desired properties for different characteristics of drugs. In this study, a chemical treatment with a widely used reagent of glutaraldehyde (GA) was selected to modify the characteristics of collagen isolated from pork skin supplied by the local slaughter house. Viscosity change with the addition of different GA concentration(0%，0.05%，0.075%，0.1%，0.2%，0.25%，0.3%) at a fixed concentration of collagen were measured by Brookfield Viscometer to examine the influence on the crosslinking rate and degree of collagen. The effects of adding different Cremopher RH40 concentration (0%，1%，5%，10%) or 1% of different active drugs (Vitamin A-Palmitate、C-Palmitate、E-Acetate、A-Acid) were examined as well. Finally, characteristics of collagen films with different extent of crosslinking collagen films (0%，0.05%，0.075%，0.1%，0.2%，0.25%，and 0.3%) were assessed using solution or gel type of drugs (the same before) with a flow through diffusion system. Results showed that the collagen viscosity increased with increasing the GA concentration. When the GA concentration was above 0.2%, the collagen viscosity reached plateur and become constant. Nevertheless, the collagen viscosity decreased with increasing Cremophor RH40 concentration, whereas active drugs didn’t show any effect on the collagen viscosity. Results of diffusion studies showed that when the GA concentration used to crosslink collagen films increased, the flux of all drugs examined decreased correspondingly. However, the flux tended to became constant When GA concentration was above 0.2%. But Vitamin A-Acid either in solution or gel form showed that when GA concentration was above 0.2%，the flux increased instead. This result was not explainable by the difference of partition coefficient measured.

並列關鍵字

SPF(Sun Protection Factor) ； Sunscreens ； Collagen ； Cross-linking agent ； Glutaraldehyde(GA)

參考文獻

First Part :1. M.A. Pathak:Sunscreens-progress and perspectives on photoprotection of human skin against UVB and UVA radiation， The Journal of Dermatology，1996，23:783-800

2. M.A. Pathak:Sunscreens and their use in the preventive treatment of sunlight-induced skin damage，The Journal of Dermatologic Surgery and Oncology，1987，13:739-750

4. D.L. Bissett，R. Chatterjee，D.P. Hannon:Photoprotective effect of superoxide-scavenging antioxidants against ultraviolet radiation-induced chronic skin damage in the hairless mouse，Photodermatology,Photoimmunology and Photomedicine，1992，7:56-62

5. F. Dreher，B. Gabard，D.A. Schwindt and H.I. Maibach: Topical melatonin in combination with vitamins E and C protects skin from ultraviolet-induced erythema:a human study in vivo，British Journal of Dermatology，1998，139: 332-339

6. A.M. Young and G. Gregoriadis:Photolysis of retinol in liposomes and its protection with tocopherol and oxybenzone， Photochemistry and Photobiology，1996，63(3):344-352